Autophagy is the segregation and delivery of cytoplasmic cargo, including proteins and organelles, for degradation by hydrolytic enzymes through the lysosomal machinery. It is critical to the healthy functioning of cells, and the failure of autophagy is a primary driver of the accumulation of cell damage and aging.

Tools for monitoring p62 and LC3B allow researchers to explore the connection between the ubitiquin degradation machinery and autophagy. We provide a range of Molecular Probes® products to investigate autophagic activity using both imaging and microplate platforms. Molecular Probes® products can be used to track lysosome incorporation into the autophagosome, and monitor proteins from synthesis through the autophagic pathway to aggregation and ubiquitination.

The Process

Schematic depiction of the autophagy pathway including p62 and LC3B. The first step involves the formation and elongation of the isolation membranes or phagophore. The second step entails the expansion and sequestering of the cytoplasm and formation of the double-membrane autophagosome. During this step the LC3 protein is recruited to the auophagosome and p62 along with the attached ubiquitin cargo attach to the LC3B protein. The fusion of lysosome with autophagosome generate the autolysosome. At this step the LC3B disassociates and the p62 and the attached ubquitin cargo are degraded.